CpG和ICTYOLANE^(TM)31佐剂对ASFV-CD2v蛋白免疫效果的影响  

Effects of CpG and ICTYOLANE^(TM)31 adjuvant on immune effect of ASFV-CD2v protein

作　　者：庞忠宝 黄颖 冯春燕[2] 翟文竹 吉日木图 朱鸿飞[1] 贾红[1] PANG Zhongbao;HUANG Ying;FENG Chunyan;ZHAI Wenzhu;JIRI Mutu;ZHU Hongfei;JIA Hong(Beijing Institute of Animal Sciences and Veterinary Medicine,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Sanya Biosafety Center of Chinese Academy of Inspection and Quarantine,Beijing 100179,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]中国检验检疫科学研究院三亚生物安全中心,北京100179

出　　处：《中国兽医学报》2023年第12期2418-2424,共7页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2021YFD1801200);云南省重点研发计划资助项目(202103AC100001);中国农业技术支持专项资金资助项目(CARS36);农业科技创新计划资助项目(ASTIP-IAS-11)。

摘　　要：探讨CpG寡聚脱氧核苷酸(CpG oligodeoxynucleotides,CpG ODN)和ICTYOLANE^(TM)31微乳佐剂对重组表达的ASFV-CD2v蛋白在小鼠体内免疫效果的影响。以杆状病毒表达的CD2v蛋白为抗原,分别配伍CpG寡脱氧核苷酸(CpG oligonucleotide,CPG ODN)和ICTYOLANE^(TM)31微乳佐剂,皮下免疫BALB/c小鼠。首次免疫后14 d进行二免,采集首免后0、14 d以及二免后7、14及28 d血清,通过间接ELISA检测血清中CD2v抗体水平;采集二免后35 d小鼠分离脾脏淋巴细胞,通过流式细胞术分析IFN-γ水平。结果显示,间接ELISA结果显示在二免后7 d可检测到较高水平的CD2v抗体,ICTYOLANE^(TM)31佐剂和CPG ODN佐剂组抗体水平明显高于蛋白组。T细胞亚群分析结果显示,在二免后35 d,CpG ODN佐剂组的CD4^(+)T淋巴细胞的比例最高,达到81.34%,ICTYOLANE^(TM)31佐剂组CD8^(+)T淋巴细胞的比例最低,仅为11.37%。ICTYOLANE^(TM)31佐剂组CD4^(+)/CD8^(+)比值最高,ICTYOLANE^(TM)31佐剂组CD4^(+)T细胞IFN-γ分泌水平最高,可达9.07%,但CpG ODN佐剂组、ICTYOLANE^(TM)31佐剂组及蛋白组间差异不显著。推断,小鼠在第二次免疫后7 d产生了较高水平的抗体,CpG ODN、ICTYOLANE^(TM)31佐剂对ASFV-CD2v蛋白均可增强免疫小鼠产生CD2v抗体的水平,其中CpG ODN佐剂作用更强。ICTYOLANE^(TM)31佐剂及CpG ODN佐剂可以增强机体细胞免疫,其中ICTYOLANE^(TM)31佐剂作用更强。该研究为非洲猪瘟亚单位疫苗相关佐剂的筛选提供了一定参考。To investigate the effects of CpG oligodeoxynucleotides(CpG ODN)and ICTYOLANE^(TM)31 microemulsion adjuvant on the immune effect of recombinant ASFV-CD2v protein in mice,CD2v protein expressed by baculovirus was used as antigen,and combined with CPG ODN and ICTYOLANE^(TM)31 microemulsion adjuvant respectively,then BALB/c mice were subcutaneously immunized.The second immunization was performed 14 d after the first immunization,and the serum was collected at 0 and 14 d after the first immunization and 7,14 and 28 d after the second immunization,and the level of CD2v antibody in the serum was detected by indirect ELISA.Splenic lymphocytes were isolated from mice 35 d after second immunization,and the level of IFN-γ secreted by CD4^(+)cells was analyzed by flow cytometry.The results of indirect ELISA showed that a high level of CD2v antibody could be detected on the 7th day after second immunization,and the antibody level in ICTYOLANE^(TM)31 adjuvant group and CPG ODN adjuvant group was significantly higher than that in protein group.T cell subgroup analysis showed that 35 d after second immunization,the proportion of CD4^(+)T lymphocytes in the CPG ODN adjuvant group was the highest,reaching 81.34%.The proportion of CD8^(+)T lymphocytes in the ICTYOLANE^(TM)31 adjuvant group was the lowest,which was only 11.37%.The ratio of CD4^(+)/CD8^(+)in ICTYOLANE^(TM)31 adjuvant group was the highest,and the level of IFN-y secretion of CD4^(+)T cells in ICTYOLANE^(TM)31 adjuvant group was the highest,up to 9.07%,but there was no significant difference among CpG ODN adjuvant group,ICTYOLANE^(TM)31 adjuvant group and protein group.Seven days after the second immunization,mice produced a high level of antibody.CpG ODN and ICTYOLANE^(TM)31 adjuvants could enhance the level of CD2v antibody in immunized mice with ASFV-CD2v protein,and the effect of CpG ODN adjuvant was stronger.In conclusion,ICTYOLANE^(TM)31 adjuvant and CpG ODN adjuvant can enhance cellular immunity,and the effect of ICTYOLANE^(TM)31adjuvant is stronger.This study

关 键 词：CpG佐剂 非洲猪瘟 CD2v 免疫抗体 IFN-Γ ICTYOLANETM31 

分 类 号：S852.65[农业科学—基础兽医学]